Biochemical and structural analyses of a higher plant photosystem II supercomplex of a photosystem I-less mutant of barley. Consequences of a chronic over-reduction of the plastoquinone pool

Photosystem II of higher plants is a multisubunit transmembrane complex composed of a core moiety and an extensive peripheral antenna system. The number of antenna polypeptides per core complex is modulated following environmental conditions in order to optimize photosynthetic performance. In this study, we used a barley (Hordeum vulgare) mutant, viridis zb63, which lacks photosystem I, to mimic extreme and chronic overexcitation of photosystem II. The mutation was shown to reduce the photosystem II antenna to a minimal size of about 100 chlorophylls per photosystem II reaction centre, which was not further reducible. The minimal photosystem II unit was analysed by biochemical methods and by electron microscopy, and found to consist of a dimeric photosystem II reaction centre core surrounded by monomeric Lhcb4 (chlorophyll protein 29), Lhcb5 (chlorophyll protein 26) and trimeric light-harvesting complex II antenna proteins. This minimal photosystem II unit forms arrays in vivo, possibly to increase the efficiency of energy distribution and provide photoprotection. In wild-type plants, an additional antenna protein, chlorophyll protein 24 (Lhcb6), which is not expressed in viridis zb63, is proposed to associate to this minimal unit and stabilize larger antenna systems when needed. The analysis of the mutant also revealed the presence of two distinct signalling pathways activated by excess light absorbed by photosystem II: one, dependent on the redox state of the electron transport chain, is involved in the regulation of antenna size, and the second, more directly linked to the level of photoinhibitory stress perceived by the cell, participates in regulating carotenoid biosynthesis.     

Attributional and Consequential Environmental Assessment of the Shift to Lead-Free Solders (10 pp)

- Goal, Scope, Background. As of July 1st, 2006, lead will be banned in most solder pastes used in the electronics industry. This has called for environmental evaluation of alternatives to tin-lead solders. Our life cycle assessment (LCA) has two aims: (i) to compare attributional and consequential LCA methodologies, and (ii) to compare a SnPb solder (62% tin, 36% lead, 2% silver) to a Pb-free solder (95.5% tin, 3.8% silver, 0.7% copper). Methods An attributional LCA model describes the environmental impact of the solder life cycle. Ideally, it should include average data on each unit process within the life cycle. The model does not include unit processes other than those of the life cycle investigated, but significant cut-offs within the life cycle can be avoided through the use of environmentally expanded input-output tables. A consequential LCA model includes unit processes that are significantly affected irrespective of whether they are within or outside the life cycle. Ideally, it should include marginal data on bulk production processes in the background system. Our consequential LCA model includes economic partial equilibrium models of the lead and scrap lead markets. However, both our LCA models are based on data from the literature or from individual production sites. The partial equilibrium models are based on assumptions. The life cycle impact assessment is restricted to global warming potential (GWP). Results and Discussion The attributional LCA demonstrates the obvious fact that the shift from SnPb to Pb-free solder means that lead is more or less eliminated from the solder life cycle. The attributional LCA results also indicate that the Pb-free option contributes 10% more to the GWP than SnPb. Despite the poor quality of the data, the consequential LCA demonstrates that, when lead use is eliminated from the solder life cycle, the effect is partly offset by increased lead use in batteries and other products. This shift can contribute to environmental improvement because lead emissions are likely to be greatly reduced, while batteries can contribute to reducing GWP, thereby offsetting part of the GWP increase in the solder life cycle. Conclusions The shift from SnPb to Pb-free solder is likely to result in reduced lead emissions and increased GWP. Attributional and consequential LCAs yield complementary knowledge on the consequences of this shift in solder pastes. At present, consequential LCA is hampered by the lack of readily available marginal data and the lack of input data to economic partial equilibrium models. However, when the input to a consequential LCA model is in the form of quantitative assumptions based on a semi-qualitative discussion, the model can still generate new knowledge. Recommendations and Outlook Experts on partial equilibrium models should be involved in consequential LCA modeling in order to improve the input data on price elasticity, marginal production, and marginal consumption.     

DNA methylation pattern in pig in vivo produced embryos

DNA methylation/demethylation pattern, determined by 5-methylcytosine (5-MeC) immunostaining, was evaluated in porcine “in vivo” produced embryos from zygote up to the blastocyst stage. In one-cell stage embryos, only the maternal pronucleus showed a positive labeling whilst the paternal pronucleus showed almost no labeling. The intensity of labeling is high until the late morula stage. Blastocysts containing less than 100 cells showed the same intensity of labeling in both the inner cell mass (ICM) nuclei and the trophectodermal (TE) cell nuclei. Interestingly, with further cell multiplication, cells of the ICM became more intensively labeled when compared to TE cells. This distinct methylation pattern is even more profound in blastocysts containing about 200–300 cells and is not caused by the difference in the cell volume of ICM and TE cells.An erratum to this article can be found at     

Suppression of HIV-1 protease inhibitor resistance by phosphonate-mediated solvent anchoring

The introduction of human immunodeficiency virus type 1 (HIV-1) protease inhibitors (PIs) markedly improved the clinical outcome and control of HIV-1 infection. However, cross-resistance among PIs due to a wide spectrum of mutations in viral protease is a major factor limiting their broader clinical use. Here we report on the suppression of PI resistance using a covalent attachment of a phosphonic acid motif to a peptidomimetic inhibitor scaffold. The resulting phosphonate analogs maintain high binding affinity to HIV-1 protease, potent antiretroviral activity, and unlike the parent molecules, display no loss of potency against a panel of clinically important PI-resistant HIV-1 strains. As shown by crystallographic analysis, the phosphonate moiety is highly exposed to solvent with no discernable interactions with any of the enzyme active site or surface residues. We term this effect "solvent anchoring" and demonstrate that it is driven by a favorable change in the inhibitor binding entropy upon the interaction with mutant enzymes. This type of thermodynamic behavior, which was not found with the parent scaffold fully buried in the enzyme active site, is a result of the increased degeneracy of inhibitor binding states, allowing effective molecular adaptation to the expanded cavity volume of mutant proteases. This strategy, which is applicable to various PI scaffolds, should facilitate the design of novel PIs and potentially other antiviral therapeutics.